Rational design and theoretical optimization of a cryopreservation protocol

Jens O.M. Karlsson, Ali Eroglu, Thomas L. Toth, Ernest G. Cravalho, Mehmet Toner

Research output: Contribution to journalConference articlepeer-review

6 Scopus citations


A cryopreservation protocol for mouse oocytes was developed by rational design and theoretical optimization using a physicochemical model of the intracellular ice formation process. Theoretical models of freeze-induced cell dehydration, homogeneous and heterogeneous nucleation of intracellular ice, and diffusion-limited crystal growth of intracellular ice were coupled and solved numerically to obtain predictions of cell response to freezing. A two-step, piecewise linear cooling protocol was chosen based on simulation results, and protocol parameters were then optimized using the mathematical model. The predicted optimal parameters were an initial cooling rate of Bdehydration = 0.59 °C/min and a plunge temperature of Tplunge = -67 °C. Experimental testing showed that optimal survival (78% cells morphologically normal) was obtained at Bdehydration = 0.5 °C/min and Tplunge = -80 °C. The fertilization rate and blastocyst formation rate of oocytes cryopreserved using the optimized protocol were 65% and 49%, respectively.

Original languageEnglish (US)
Pages (from-to)85-89
Number of pages5
JournalAmerican Society of Mechanical Engineers, Heat Transfer Division, (Publication) HTD
StatePublished - Dec 1 1995
EventProceedings of the 1995 ASME International Mechanical Engineering Congress and Exposition - San Francisco, CA, USA
Duration: Nov 12 1995Nov 17 1995

ASJC Scopus subject areas

  • Mechanical Engineering
  • Fluid Flow and Transfer Processes


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